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Current Reviews in Clinical and Experimental Pharmacology, 2021, 16, 73-90 73

RESEARCH ARTICLE

Formulation and Evaluation of Polyherbal Cream and Lotion for the

Treatment of Psoriasis-Induced Secondary Infections

Priyanka Joshi1, Sushil Joshi1,*, Urvashi Rajani2, Ruchi B. Semwal3 and Deepak K. Semwal4,*

1
Research and Development, Patanjali Ayurved Ltd, Haridwar-249402, Uttarakhand, India; 2Saurashtra University,
Rajkot-365601, Gujarat, India; 3Department of Chemistry, Pt. L.M.S. Govt. PG College, Rishikesh, Uttarakhand, India;
4
Department of Phytochemistry, Faculty of Biomedical Sciences, Uttarakhand Ayurved University, Dehradun-248001,
Uttarakhand, India

ARTICLE HISTORY
Received: April 21, 2019
Revised: September 16, 2019
Accepted: September 25, 2019
DOI:
10.2174/1574884714666191017111218
Keywords: Malassezia pachydermatis, essential oil, eugenol, Candida spp, scalp psoriasis, amplification.
Abstract: Background: Psoriasis is one of the most common skin diseases in humans and affects a
major population worldwide. The aim of the present study is to evaluate the efficacy of selected
polyherbal formulations against psoriasis-induced secondary infections.
Aim: Psoriasis is one of the most common skin diseases in humans and affects a major population
worldwide. The aim of the present study is to evaluate the efficacy of selected polyherbal formula-
tions against psoriasis-induced secondary infections.
Methods: Samples were collected from the scalp, behind the ears, chest and back of the patients.
The microscopic examination of fungal and bacterial growth was carried out with lactophenol cot-
ton blue stain and gram stain, respectively. Volatile constituents of essential oils were identified by
GC-MS analysis, in order to investigate the relation between chemical composition and biological
activity. Nutrient agar media was used for antibacterial activity whereas Sabourauds dextrose agar
media was used for antifungal activity.
Results: A total of 24 isolates were obtained from 2 patients of scalp psoriasis. Anti-oxidant activity of
the samples was determined using DPPH radical scavenging method. In the present study, EOs
showed a good antifungal action against Malassezia pachydermatis and other microbial strains. The
GC-MS analysis revealed the presence of eugenol, linalool, citral, neral, limonene, terpenes, eucalyp-
tol and thymol in the essential oils. The formulated retention lotion 1 (L4), 2 (L5), 3 (L6) and cream 1
(C1) showed DPPH radical scavenging activity by 23.52%, 24.48%, 28% and 5.08%, respectively.
Conclusion: The present study concluded that most of the formulated lotions and creams showed
good antimicrobial activity and may be applied topically against scalp psoriasis.

1. INTRODUCTION
Psoriasis is a common inflammatory and chronic skin
disease affecting both men and women. Its prevalence in
India ranges from 0.44 to 2.8%. The proportion of psoriasis
was found to be 2.6% out of all skin diseases [1]. A person’s
genetic makeup can also be the reason for the development
of this disease [2]. Its beginning involves T cells (a type of
WBC), immune system sends such signals that put T cells in
action and become overly active, which leads to unhealthy
*Address correspondence to these authors at the Research and Development,
Patanjali Ayurved Ltd, Haridwar-249402, Uttarakhand, India;
E-mail: sushil.joshi@patanjaliayurved.org and Department of Phytochemis-
try, Faculty of Biomedical Sciences, Uttarakhand Ayurved University, De-
hradun-248001, Uttarakhand, India; Tel: +91-135-2685124;
E-mail: dr_dks.1983@yahoo.co.in
swelling and fast turnover of skin cells. Although any part of
the body can be affected but knees, elbows, knuckles, lower
back and sacral areas are most commonly affected [3].
Breaking, pitting, thickening of the nail or thickening under
the nails are the symptoms of Nail psoriasis which is the
rarest type of Psoriasis. Around 20% of people with psoriasis
are suffering from psoriatic arthritis which is very painful
[4]. It is an incurable disease, only symptomatical relief can
be achieved and severity can be lessened. Symptoms can be
cleared for months or even years but it is a lifelong disease
[5]. Affected skin is itchy and irritating, people suffering
from psoriasis may also suffer from Type-2 diabetes, in-
flammatory bowel disease, cardiovascular disease, psoriatic
arthritis and microbial skin infections. On the basis of sever-
ity, location, duration, shape, size and pattern of scales, pso-
riasis can be classified into the following types: Pustular,

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74 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1
Erythrodermic, Inverse, Guttate, Plaque, Psoriatic, Nail and
Scalp Psoriasis.
Researchers around the world have identified several
areas on the human genome where more than one gene may
be involved in psoriasis and psoriatic arthritis [6]. More re-
cent studies have suggested multiple loci on many different
chromosomes besides chromosome 6, including chromo-
somes 1, 2, 4, 8, 16, 5, and 20 [7-14]. While there seem to be
multiple genes that contribute to the psoriatic phenotype,
most investigators focus on the MHC I region, with particu-
lar interest on the HLA-Cw6 allele [15]. Localization of a
second psoriasis susceptibility locus (PSORS2) to chromo-
some 17q25 was achieved following a genome-wide linkage
scan on eight multiply affected families [16].
With the employment of genome-wide scans, investiga-
tors have been able to map (with varying degrees of confi-
dence) at least six different susceptibility loci, designated
PSORS1–PSORS6 [15]. Several other psoriasis liable loci
have also been mapped, including PSORS7 (1p) and
PSORS9 (4q31), and additional studies are ongoing in many
laboratories [16-18]. Studies have reported the foremost ge-
netic determining factor for psoriasis that is within the
PSORS1 region of the MHC on chromosome 6p21 [19]. The
two most likely candidate loci at 6p21.3 are HLA-Cw*0602
and the Corneodesmosin (CDSN) gene that encode an adhe-
sive protein expressed by keratinocytes and is important in
terminal differentiation of the epidermis. Thus, it is predict-
able that genes such as HLA-Cw6 are in linkage disequilib-
rium with another gene, or block of genes, at a distinct locus
such as Killer Ig-like Receptor (KIR) [20]. The second most
well-characterized disease-susceptibility locus (PSORS2)
resides within 17q24–q25 [21]. A susceptibility locus for
psoriasis, PSORS4, has been mapped to chromosome 1q21
in the region of the epidermal differentiation complex [19].
Psoriasis is a polygenic, chronic relapsing inflammatory
autoimmune disease based on the interplay between antigen-
presenting dendritic cells, T cells and keratinocytes creating
the characteristic mark of skin lesions that are sharply de-
fined reddish and scaly plaques [22]. Particularly, myeloid
dendritic cell secretion of IL-23 and IL-12 activates IL-22,
IL-17-producing T cells, Th22 and Th1 cells, resulting in the
production of inflammatory cytokines such as IL-17, 23IFN-
γ, TNF and IL-22. These cytokines facilitate effects on
keratinocytes thus creating the inflammatory loop [23].
The immunopathogenesis of psoriasis is complex and
involves variations in the innate immunologic system
(keratinocytes, dendritic cells, histiocytes, neutrocytes, mas-
tocytes, endothelial cells) and acquired (T lymphocytes)
[25]. When the cells of the Innate Immunologic System (IIS)
are activated, they produce growth factors, cytokines and
chemokines that act upon the cells of the Acquired Immu-
nologic System (AIS) and vice-versa [24-28].
The current view of psoriasis pathogenesis points to-
wards the fact that the aberrant immune and epidermal re-
sponse was seen in psoriasis is sustained by a pathogenic
cross-talk between epithelial and immune cells [29, 30]. This
interplay is mainly compelled by the critical pro-
inflammatory molecules, TNF, IL-23 and IL-17, whose di-
rect therapeutic targeting has proven to be clinically effi-
Joshi et al.
cient, with other mediators, such as IFN-α, IFN-γ and IL-22
possibly contributing to the initiation, amplification and
maintenance of the disease [31].
An extensive literature also has suggested that bacteria,
including Staphylococcus aureus [32] and Streptococcus
pyogenes [33], could play a role in the induction and mainte-
nance of psoriasis, and Group A Streptococcus (GAS) anti-
gens or superantigens have been implicated in psoriasis
pathogenesis in genetically predisposed individuals [34, 35].
The most abundant and diverse phylum populating the
psoriatic lesions is Firmicutes (46.2%) followed by Actino-
bacteria (37.3%). However, Actinobacteria is the most
prevalent and diverse phylum in the samples from the nor-
mal skin of both healthy persons (47.6%) and from the pa-
tients with psoriasis (47.8%). Proteo bacteria are found less
frequently in psoriatic lesion samples (11.4%), compared
with samples from healthy persons [36].
Among these phyla, Coryne bacterium, Staphylococcus,
Streptococcus, and Propioni bacterium are the dominant
genera in the normal skin, including healthy persons and
patients with psoriasis, as well as in the psoriatic lesion. The
most common genera detected in the samples from normal
and diseased skin were similar, but the proportions differed.
The ratio of Streptococcus to Propioni bacterium (S/P ratio)
in healthy persons and normal skin of patients with psoriasis
is 0.4. In contrast, the S/P ratio (5.0) in the psoriatic lesions
is significantly higher [37].
Fungal microbiota has also been involved in the aetiol-
ogy of psoriasis; Malassezia organisms have been implicated
in both scalp and facial psoriasis. Furthermore, Candida al-
bicans is frequently found in the case of psoriasis [38, 39].
Five Malassezia species and four unknown phylotypes are
identified. Malassezia restricta is the commonest species
found, but Malassezia globosa is also frequently detected
[40].
All nine of Malassezia species are detected at different
rates in patients with psoriasis. The overall detection rates in
lesional and non-lesional skin of M. restricta, M. globosa
and M. sympodialis are high (96%, 82% and 64%, respec-
tively), whereas the detection rates of the other species are
relatively low. However, there is no difference in the rates
between lesional and non-lesional skin areas [41].
M. restricta and M. globosa account for more than70% of
all Malassezia sequences in both groups; however, the ratio
of these two species differs between the groups. M. restricta
levels are significantly higher in the psoriatic group, com-
pared with controls whereas M. globosa levels are higher in
the control group [42]. Candida spp. are found in 15 % of
skin specimens from patients with psoriasis compared with 4
% of controls (p = 0.045), and in oral specimens of 60 % of
patients with psoriasis compared with 20 % of controls [43].
Many of the molecular pathways linked with psoriasis
pathogenesis are also involved in host defence mechanisms
that protect against common pathogens. Candida can arouse
the production of cytokines that trigger or exacerbate psoria-
sis, and many systemic psoriasis treatments may put patients
at increased risk for developing oral, cutaneous, and genitou-
rinary candidiasis. Therefore, dermatologists should regu-
Polyherbal Cream and Lotion to Treat Psoriasis Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 75
larly monitor patients with psoriasis for signs of Candida
infection, and take steps to successfully treat these infections
to avert worsening of psoriasis symptoms [44].
The demand for herbal or Ayurvedic formulations has
increased significantly in the last few years. People encoun-
ter a number of skin and hair disorders due to the modern
world’s changing food habits and increased stress levels [45,
46]. The poor environmental conditions can also be attrib-
uted to rising cosmetic issues. Since these factors are un-
avoidable, therefore there is a need for a treatment that could
counter all the problems in a much safer way [47]. The anti-
fungal, as well as conditioning properties of the plant ex-
tracts, make them an effective alternative to chemical agents
in various natural anti-microbial formulations [48].
Present study mainly focuses on the evaluation of effi-
cacy of various plant extracts such as Psidium guajava, Law-
sonia inermis, Phyllanthus emblica, Syzygiuma romaticum,
Acacia concinna, Piper nigrum, Juglans regia, Sapindus
mukorossi, Salvador apersica, Ociumum tenuiflorum,
Trigonella foenum-graecum, Curcuma longa, Aloe vera,
Withania somnifera and essential oils against the isolated
microorganisms from scalp and lesions of patients and for-
mulate polyherbal cream and lotion for scalp psoriasis.
2. MATERIALS AND METHODS
2.1. Collection and Microscopic Examination of Clinical
Samples
Samples were collected from mid-region of the hair, be-
hind the ears, chest and back area of the two patients for the
isolation of microorganisms from psoriatic lesions. Sterile
cotton swabs were soaked in the tubes containing peptone
water and Sabourauds Dextrose Broth (SDB), respectively
and each of the two swabs was rubbed along each of the
marked areas such as to cover the total surface in a non-
overlapping manner. Cotton swabs with no skin contact
submitted to the same procedures were used as negative con-
trols [49].
For identification, isolates samples collected in SDB
were incubated at 25oC for 5 days and then cultured on SDB
incorporated with chloramphenicol to avoid bacterial con-
taminants. The plates were incubated at room temperature
(25oC) for three days and then, up to a week. The isolates in
peptone water were incubated at 37oC for 48 hours and then
cultured on Nutrient agar plates. The plates were again incu-
bated at 37oC for 48 hours to carry out further identification.
The microscopic examination of fungal and bacterial growth
was carried out with lactophenol cotton blue stain and gram
stain respectively.
2.2. Biochemical Characterization
The bacterial isolates were characterized biochemically
by Simmons citrate test, urease test, catalase test, coagulase
test, indole test, H2S production, Starch hydrolysis test and
sugar fermentation tests [50].
2.3. Composition of Phenol Red Carbohydrate Broth
Phenol red carbohydrate broth comprised of trypticase or
protease peptone No. 3 (10 g), sodium chloride (5 g), phenol
red (7.2 ml of 0.25% phenol red solution) (0.018 g) and car-
bohydrate source (10 g). The broth medium was prepared by
mixing all the ingredients in 1000 ml of distilled water. 4-5
ml of this broth was filled in test tubes. Inverted Durham
tubes were inserted to detect gas production. The prepared
test media was sterilized by autoclaving at 121ºC for 15
minutes. All the test tubes were inoculated with the test mi-
croorganism. One tube was kept as control. The tubes were
incubated at 37ºC for 24 hours and then observed for
changes [51]. A change in the colour of media from red to
yellow indicated a positive result for acid production. The
formation of a bubble in the Durham tube illustrated a posi-
tive result for gas production.
2.4. Collection of Plant Material
The raw material for extracts includes Psidium guajava,
Lawsonia inermis, Phyllanthus emblica, Syzygiuma romati-
cum, Acacia concinna, Piper nigrum, Juglans regia, Sapin-
dus mukorossi, Salvador apersica, Ociumum tenuiflorum,
Trigonella foenum-graecum, Curcuma longa, Aloe vera, and
Withania somnifera. These plants were collected from the
local market and shade dried prior extraction. The Essential
Oils (EO) required for testing and formulation of the product
were obtained from the store of Patanjali Natural Coloroma
Pvt. Ltd.
2.5. Preparation of Extracts
Aqueous extracts were prepared for all the raw materials.
50g of dried and powdered sample was filled in a round bot-
tom flask and refluxed with 100 ml of water. The tempera-
ture was maintained at 100°C. All the extracts were filtered
through a Whatman filter paper No. 1 and then concentrated
by using a rotary evaporator 40-50°C under vacuum. The
dried extracts were collected and stored under dark under
refrigerated conditions [52].
2.6. Phytochemical Analysis of Extracts
Chemical tests for saponins, anthocyanins, flavonoids,
alkaloids, tannins and phenolic compounds were conducted
by standard protocols [52]. Preliminary phytochemical
analysis was carried out for the extracts as per standard
methods described by Brain and Turner (1975) and Evans
(1996).
2.7. Quantitative Analysis of Essential Oils
The volatile constituents of each EO were analyzed by
GC-MS. It was accomplished with a Hewett-Packed gas
chromatographer (HP 6890) coupled with a mass spectro-
photometer. Fragmentation was performed by electron im-
pact at 70ev. The column used was HP 5MS (30m 0.25µm).
The carrier gas is helium with a flow rate of 1.5ml min-1. The
injection mode was split (split ratio: 1/70, flow rate 64.2ml
min-1). The column temperature was programmed from 60 to
2200C at a heating rate of 40Cmin-1, for 5 min. The purge
flow is 3.0ml/min and linear velocity is 40.ocm/sec. for the
chromatographic analysis. The essential oil was diluted in
methanol (1/10 v/v). The injection and detector temperature
were 250 and 2800C. The identification of the components is
based on the comparison of their mass spectra (GC/MS),
respective with spectra [53].
76 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1
Identification of the constituents was based on a com-
parison of the retention time with those of authentic samples,
comparing their linear indices relative to a series of n alkanes
(C8-C23). Further identifications were also made possible by
the use of a homemade library of mass spectra built up from
pure substances and components of known oils, and MS lit-
erature data (NIST 2000, ADAMS) [53].
2.8. Formulation of Product
One cream, two lotions and three retention lotion were
formulated and evaluated for antimicrobial activity. Each
sample contains a different set of active ingredients. A list of
all the formulations is given below. Out of these four best
formulations were selected on the basis of the highest activ-
ity and were taken into account for further evaluation and
trials.
2.9. Qualitative Analysis by Thin-layer Chromatography
2.9.1. TLC of Extracts and Products
The above-prepared plant extracts were diluted in metha-
nol (50 mg/ml) and formulations were diluted in methanol at
10mg/ml concentration for TLC application. The prepared
dilutions were applied on pre-coated TLC plates by using
capillary tubes and developed in a chamber using a suitable
mobile phase (5 toluene: 4 ethyl acetate: 1 formic acid). The
developed TLC plates were air-dried and sprayed with ani-
saldehyde-sulphuric acid spraying reagent and were placed
in a hot air oven for 1 min for the development of coloured
bands [54].
2.10. Quantitative Analysis by HPLC
Extracts were standardized using HPLC. The system
consisted of variant pumps with a three-pump gradient sys-
tem-model combined with a fixed wavelength UV-VIS de-
tector, a sample injector with 20µl sample loop. The column
was a C18 (25cm long x 4.6 mm dia) and 5 µm particle di-
ameter. Detection was carried out by measurement of UV
absorbance at 272 nm. The mobile phase was composed of
water: methanol (95:5v/v), pH 3 adjusted by orthophosphoric
acid. The mobile phase flow was maintained at 1 ml/min.
The percentage of standard (ascorbic acid) in extract and
product was calculated using the formula, % of std in sample
= (sample area/std area) × (std dilution/sample dilu-
tion)×purity of the std.
2.11. Determination of the Antioxidant Activity of Prod-
uct with DPPH Radical Scavenging Method
The samples were evaluated for antioxidant activity by
DPPH scavenging method. The DPPH radical absorbs at 517
nm, but its absorption decreases upon reduction by an anti-
oxidant or a radical. Briefly, 0.1 mM solution of DPPH was
prepared in ethyl alcohol and 1 mL of this solution was
added to 800 µl of tris HCl buffer and 200µl of extract solu-
tion in phosphate buffer of concentration (1 mg/mL). These
solutions were vortexed thoroughly and incubated in dark for
30 min. Thirty minutes later, the absorbance was measured
at 517 nm against blank samples lacking scavenger [55]. The
percent DPPH scavenging effect was calculated using the
following equation:
Joshi et al.
DPPH scavenging effect = {(A0 – A1)/A1} × 100
Where A0 is the absorbance of the control reaction and
A1 is the absorbance in the presence of extract [56].
2.12. Determination of Biological Activity
2.12.1. Biological Activity of Extracts, Essential Oils and
Products
To check the antimicrobial efficacy of ingredients and
products, different concentrations of aqueous plant extracts
were made in water and were autoclaved before testing. All
the essential oils were diluted in DMSO making a concentra-
tion of 1%. For the products, all the lotion and cream were
diluted in DMSO in the ratio 1:4. The antimicrobial activities
were evaluated using Agar Well Diffusion method [57].
2.13. Antibacterial Activity
For evaluation of the antibacterial activity, Nutrient agar
media was sterilised and poured into sterile Petri plates and
left to solidify. 30µL of bacterial culture was spread on the
solidified media using a sterilised glass spreader. A few
minutes later, a well was made in the media using a cork
borer and 300µL of the sample was filled in the well. The
Petri plates were then incubated at 37ºC for 48 hours. After
incubation, the zone of inhibition of each extract was meas-
ured using the antibiotic zone scale.
2.14. Antifungal Activity
For evaluation of antifungal activity, Sabourauds dex-
trose agar media was sterilized and poured into sterile petri
plates and left to solidify. 30µL of fungal culture was spread
on the solidified media using a sterilized glass spreader. A
few minutes later, a well was made in the media using a cork
borer and 300µL of the sample was filled in the well. The
Petri plates were then incubated at 25oC for 72 hours. After
incubation, the zone of inhibition of each extract was meas-
ured using the antifungal zone scale.
3. RESULT AND DISCUSSION
Skin is the primary barrier against infections which plays
a vital role in the immunity of an individual. Any disruption
in the skin may enable the attack of microorganisms making
it easier for the infections to reach the internal organs. Thus,
the treatment of these lesions by using several herbal antimi-
crobial agents may provide relief to the patient.
In the present study, a total of 2 samples were collected
from the two patients with scalp psoriasis. Out of 2 samples,
a total of 24 isolates were obtained. Hence, significant con-
sideration should be given for treating the microbial infec-
tion by these fungi and bacteria. The presence of these mi-
croorganisms should be pre-screened to prevent such infec-
tions to develop and proceed at a later stage. The predomi-
nant colonies were selected and identified through colony
morphology, staining property and biochemical tests. As a
result, the organisms identified were characterized and ar-
ranged in terms of their phylogenetic relationship. The bacte-
rial strains form two major clusters i.e. bivariate dendro-
gram. This establishes the linkage within the groups of bac-
terial species (Table 1, Figs. 1-4).
Polyherbal Cream and Lotion to Treat Psoriasis Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 77

Table 1. Biochemical characterization.

Isolated Bacteria Catalase Coagulase Sulphide Indole Motility

PNCPL/PMOB-1 + - + - -

PNCPL/PMOB-2 + + - - -

PNCPL/PMOB-3 + + - - -

PNCPL/PMOB-4 + - - - +

PNCPL/PMOB-5 + - - - -

PNCPL/PMOB-6 + + - - -

PNCPL/PMOB-7 + + - - -

Isolated Bacteria Methyl Red Voges-Proskauer Urease Citrate

PNCPL/PMOB-1 + - - +

PNCPL/PMOB-2 + - - +

PNCPL/PMOB-3 + - - +

PNCPL/PMOB-4 + - - +

PNCPL/PMOB-5 + - + +

PNCPL/PMOB-6 + - - +

PNCPL/PMOB-7 - - - +

Isolated Bacteria Starch Glucose Mannitol Lactose

PNCPL/PMOB-1 - ++ ++ +

PNCPL/PMOB-2 - ++ ++ +

PNCPL/PMOB-3 - ++ ++ +

PNCPL/PMOB-4 - + ++ -

PNCPL/PMOB-5 - ++ ++ +

PNCPL/PMOB-6 - ++ ++ +

PNCPL/PMOB-7 - ++ ++ +

The present investigation shows significant variation in
the alkaloids, flavonoids, phenol and tannin content com-
pared to the standard mentioned results. These variations are
due to a number of environmental factors such as climate,
altitude, rainfall, etc. The present study showed that the plant
extracts have the potential to act as a source of useful drugs
owing to the presence of these phytochemical constituents
(Table 2). Among all the selected extracts clove has the
highest amount of flavonoids, phenolic and tannin content.
Phenols, when mixed with the flavonoid compounds in
plants, are reported to show multiple activities like antioxi-
dant, anti-carcinogenic, anti-inflammatory, etc. Tannins in-
hibit the pathogenic fungi and antimicrobial activity of ex-
tracts showed better activity by the presence of tannins. The
plant-based compounds have effective dosage response and
minimal side effects when compared to the synthetic com-
pounds.
In the present study, EOs showed a good antifungal ac-
tion against M. pachydermatis and other fungal and bacterial
strains (Tables 5, 6 and Figs. 5, 6). Their good biological
activity may be related to the chemical composition. The
EOs were characterized by the presence of eugenol, linalool,
citral, neral, limonene, terpenes, eucalyptol and thymol as
detected in GC-MS (Table 3). Even if potential antifungal
effects of some compounds from plants have recently at-
tracted attention, there is scant scientific information about
anti-Malassezia properties of herbal remedial. To the best of
our knowledge, EOs show eudermic, lenitive, antimicrobial
and anti-inflammatory activities and may contribute to skin
healing. Moreover, the use of compounds with a pleasant
smell is appreciated by the owners, fighting the strong odour
of rancid fat, characteristic of Malassezia skin disorders. For
these reasons, the choice of EOs could be an intriguing alter-
native.
Fig. 5 depicts the activity of various extracts, oils and
formulated products (lotions and cream) on each of the iso-
lated bacterial strains. Clove extract showed the highest ac-
tivity against bacterial samples whereas miswak, shikakai
78 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 Joshi et al.

Fig. (1). Pure colonies of isolated fungal strains streaked on Sabraouds Dextrose Agar medium. (A). PNCPL/PMOF-2, (B). PNCPL/PMOF-
3, (C). PNCPL/PMOF-4, (D). PNCPL/PMOF-5, (E). PNCPL/PMOF-6, (F). PNCPL/PMOF-7, (G). PNCPL/PMOF-8, (H). PNCPL/PMOF-9,
(I). PNCPL/PMOF-10, (J). PNCPL/PMOF-11, (K). PNCPL/PMOF-12, (L). PNCPL/PMOF-13, (M). PNCPL/PMOF-A, (N). PNCPL/PMOF-
B, (O). PNCPL/PMOF-D, (P). PNCPL/PMOF-F. (A higher resolution / colour version of this figure is available in the electronic copy of the
article).

extract and black pepper powder showed minimum activity
and walnut and reetha extract have no activity against bacte-
rial samples. Among the formulated products highest activity
is shown by retention lotion 3 (L6) whereas retention lotion
2 (L5) shows minimum activity. Thyme oil, cinnamon oil
and lemon grass oil exhibited the highest activity against
bacterial samples and coconut oil, neem seed oil, eucalyptus
and jojoba oil showed the minimum activity. The oil combi-
nation of lotion 2 (L2) shows the highest activity in compari-
son to other oil combinations but due to the presence of wax
and other chemicals in the lotion, the activity of the L2 de-
creases. The extracts combination of lotion 3 shows the
highest activity in comparison to other combination but due
to the presence of wax and other chemicals in lotion, the
activity of L3 decreases.
Similarly, Fig. 6 depicts the activity of various extracts,
oils and formulated products against each of the isolated
fungal strains. Shikakai and reetha extract exhibited the
Polyherbal Cream and Lotion to Treat Psoriasis Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 79

Fig. (2). Microscopic view of isolated fungal strains. (A). PNCPL/PMOF-2, (B). PNCPL/PMOF-3, (C). PNCPL/PMOF-5, (D).
PNCPL/PMOF-6, (E). PNCPL/PMOF-7, (F). PNCPL/PMOF-8, (G). PNCPL/PMOF-9, (H). PNCPL/PMOF-10, (I). PNCPL/PMOF-11, (J).
PNCPL/PMOF-12, (K). PNCPL/PMOF-13, (L). PNCPL/PMOF-A, (M). PNCPL/PMOF-B, (N). PNCPL/PMOF-D, (O). PNCPL/PMOF-F.
(A higher resolution / colour version of this figure is available in the electronic copy of the article).

highest activity whereas clove extract, walnut extract and to other combination but the presence of wax and other
black pepper powder show minimum activity and miswak chemicals in lotion, the activity of the L5 decreases.
extract showed no activity. Among the formulated products
TLC profiling of all extracts gives an impressive result
highest activity was shown by retention lotion 1 (L4)
directing towards the presence of a number of phytochemi-
whereas lotion 2 (L2) showed minimum activity. Thyme oil,
cals. Various phytochemicals give different Rf values in the
cinnamon oil and lemon grass oil showed the highest activity
different solvent systems. This variation in Rf values of the
and coconut oil, neem seed oil, eucalyptus and jojoba oil
phytochemicals provides a very important clue in the under-
showed the minimum activity. The oil combination of reten-
tion lotion 1 (L4) showed the highest activity in comparison standing of their polarity. Moreover, the Rf values of prod-
ucts matched with that of the extracts showing various con-
to other oil combinations. The extracts combination of reten-
stituents of extracts being present in the products. Ascorbic
tion lotion2 (L5) showed the highest activity in comparison
80 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 Joshi et al.

Fig. (3). Biochemical tests. (A). Urease, (B). Catalase, (C). MR, (D). Citrate, (E). VP, (F). SIM, (G). Starch hydrolysis, (H). Glucose fermen-
tation, (I). Mannitol fermentation, (J). Lactose fermentation. (A higher resolution / colour version of this figure is available in the electronic
copy of the article).

acid is known to play a prominent role in scavenging free
radicals along with a contribution to organoleptic properties.
It is ubiquitously present in all fruit extracts and richly en-
dowed in herb extracts responsible for rendering antioxidant
activities and therapeutic properties. In addition to quality
enhancement, they also impart colour and flavour to the
plant extracts. The concentration of ascorbic acid was found
to be 0.0203% in shikakai extract. The experiment was per-
formed using standard HPLC protocols.
In the DPPH assay, the antioxidants were able to reduce
the stable radical DPPH to the yellow coloured diphenyl-
picrylhydrazine. DPPH is usually used as a reagent to evalu-
ate free radical scavenging activity of antioxidants. Scaveng-
ing activity of retention lotion 3 (L6) was found to be 28%.
On the other hand, retention lotion 1 (L4), retention lotion 2
(L5), lotion 2 (L2), lotion 3 (L3) and cream 1 (C1) exhibited
23.52%, 24.48%, 9.46%, 10.47% and 5.08 % DPPH radical
scavenging activity at the same concentration, respectively
(Table 4).
Aqueous extracts showed effective anti-microbial activity
against the isolated microbes. Especially clove, reetha and
shikakai plant extracts showed greater anti-microbial activity.
Polyherbal Cream and Lotion to Treat Psoriasis Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 81

Fig. (4). Dendrogram using average linkage (within groups).

Table 2. Quantitative estimation of phytochemicals in the extracts.

Sample Flavonoids Phenols Tannins

O.D. Conc. (mg/g) O.D. Conc. (mg/g) O.D. Conc. (mg/g)

Clove 0.5267 54 2.5898 85.7 1.9772 227.4

Miswak 0.2836 29.7 0.4358 13.9 0.0937 18.1

Walnut 0.2715 28.5 0.4207 13.4 0.0698 15.5

Black pepper powder 0.226 24 0.2247 6.89 0.0249 10.5

Shikakai 0.2845 29.8 - - - -

Table 3. Quantitative estimation of essential oils by GC-MS.

Compound Name RT %Area SD

Eucalyptus oil

α-Pinene 7.459 2.297 0.0058

o-Cymene 10.816 10.557 0.0208

D-Limonene 11.021 7.687 0.0416

Eucalyptol 11.194 78.647 0.0351

γ-Terpinene 12.488 0.817 0.0115

(Table 3) contd….
82 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 Joshi et al.

Compound Name RT %Area SD

Lemon grass oil

3-Carene 7.434 0.337 0.025

Camphene 7.925 1.793 0.098

5-Hepten-2-one, 6-methyl- 9.085 1.530 0.079

2,3-Dehydro-1,8-cineole 9.349 0.127 0.012

m-cymene 10.775 0.057 0.006

D-Limonene 10.96 1.730 0.062

Eucalyptol 11.116 0.117 0.006

d-α-pinene 11.317 0.117 0.006

cis-ocimene 11.843 0.087 0.006

4-Nonanone 13.117 0.630 0.035

Terpinolene 14.112 0.097 0.006

Linalyl acetate 14.435 1.467 0.040

trans-Verbenol 15.098 0.077 0.023

2-Cyclopentene, 4- (hydroxymeth14.112yl)-1,1,2,3-14.735 Tetram15.098ethyl- 15.476 0.173 0.012

Sabinol 17.51 0.137 0.006

Cyclohexanol, 5-methyl-2-(1-methylethenyl)- 17.995 0.200 0.017

1-(1-Ethyl-2,3-dimethylcyclopent-2-enyl)-ethanone 18.336 0.270 0.010

Citronellal 18.566 1.007 0.029

endo-Borneol 19.343 0.603 0.015

Levo-menthol 19.754 0.337 0.015

2-((3,3-Dimethyloxiran-2-yl)methyl)-3-methylfuran 19.922 0.207 0.006

cis-p-Mentha-2,8-dien-1-ol 20.136 0.117 0.015

Isoneral 20.313 0.550 0.036

α-Terpineol 20.691 0.207 0.006

2,7-Octadiene-1,6-diol, 2,6-dimethyl- 21.369 0.213 0.012

Citronellol 22.273 2.267 0.0950

Citral 22.803 24.697 0.170

Geraniol 23.226 12.577 0.114

Neral 23.793 33.820 0.046

Geranyl vinyl ether 23.999 0.217 0.012

β-Pinene 24.611 0.447 0.006

6-Octen-1-ol, 3,7-dimethyl-, acetate 25.877 0.643 0.006

α-ylangene 26.399 0.143 0.006

α-Copaene 26.493 0.067 0.012

Geranyl acetate 26.571 30860 0.026

(-)-β-Bourbonene 26.715 0.067 0.006

(Table 3) contd….
Polyherbal Cream and Lotion to Treat Psoriasis Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 83

Compound Name RT %Area SD

Lemon grass oil

β-elemen 26.839 0.473 0.006

Z-β-Caryophyllene 27.467 3.017 0.032

Eugenol 27.969 0.457 0.045

(+)-epi-Bicyclosesquiphellandrene 28.885 0.227 0.012

γ-Himachalene 29.103 0.017 0.046

Naphthalene, 1,2,3,4,4a,5,6,8a-octahydro- 7-methyl-4-methylene-1-(1-methylethyl)-, (1-α,4a-β,8a-α) 29.23 1.173 1.876

Caryophyllene 30.451 2.623 1.328

Lemon peel oil

3-Carene 7.327 1.770 0.098

β-Phellandrene 8.503 1.160 0.072

β-Pinene 8.659 13.053 0.653

β-Myrcene 8.984 1.870 0.096

o-Cymene 10.299 2.730 0.128

D-Limonene 10.537 58.377 2.180

γ-Terpinene 11.671 0.890 0.035

7-Octen-2-ol, 2,6-dimethyl- 12.131 0.723 0.072

L-α-Terpineol 18.147 11.080 0.372

Cyclohexanol, 1-methyl-4-(1-methylethylidene)- 18.472 1.457 0.051

Decanal 18.862 0.720 0.338

Neral 20.855 1.150 0.030

Geranial 22.532 1.767 0.060

Bicyclo[5.2.0]nonane, 2-methylene-4,8,8-trimethyl-4-vinyl- 30.302 1.253 0.042

Cinnamon oil

Linalool 14.773 6.30 0,0058

(Z)-3-Phenylacrylaldehyde 21.923 0.10 0.0231

Cinnamaldehyde, (E)- 23.817 49.78 0.1155

Eugenol 26.04 9.36 0.0058

α-Copaene 26.492 0.06 0.0000

Bicyclo[5.2.0]nonane, 2-methylene-4,8,8-trimethyl-4-vinyl- 27.191 0.02 0.0000

Caryophyllene 27.483 11.58 0.0058

1,4,7,-Cycloundecatriene, 1,5,9,9-tetramethyl-, Z,Z,Z- 28.136 0.09 0.0058

Benzyl Benzoate 32.87 22.71 0.0981

Thyme oil

Thuzene 7.2 0.303 0.015

R-α-pinene 7.442 0.377 0.012

Camphene 7.923 0.030 0.000

(Table 3) contd….
84 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 Joshi et al.

Compound Name RT %Area SD

Thyme oil

β-Terpinene 8.885 2.507 0.081

β-Myrcene 9.275 0.363 0.015

3-Carene 10.159 0.050 0.000

α-terpinene 10.434 0.157 0.006

o-cymene 10.824 29.057 0.331

Limonene 11.022 0.540 0.010

γ-Terpinene 12.559 36.770 0.269

Benzene, 1-ethenyl-3,5-dimethyl- 14.169 0.187 0.006

Terpinen-4-ol 20.025 0.203 0.006

Benzenemethanol, α,α,4-trimethyl- 20.415 0.097 0.015

α-Terpineol 20.699 0.093 0.006

Thymol 24.368 28.897 0.423

Benzene, 1,2,3-trimethoxy-5-(2-propenyl)- 29.805 0.040 0.000

Table 4. Anti-oxidant activity of the products.

Sample Name O.D. Value % Age Inhibition

Standard 0.9797 31.00

Lotion-2 (L2) 0.8870 9.46

Lotion-3 (L3) 0.8771 10.47

Retention Lotion-1 (L4) 0.7492 23.52

Retention Lotion-2 (L5) 0.7398 24.48

Retention Lotion-3 (L6) 0.7053 28.00

Cream-1 (C1) 0.9299 5.08

Table 5. ZOI (mm) of samples against patient bacterial sample.

Sample PNCPL/PMOB-1 PNCPL/PMOB-2 PNCPL/PMOB-3 PNCPL/PMOB-4 PNCPL/PMOB-5 PNCPL/PMOB-6 PNCPL/PMOB-7

Lotion 2 (20 %) 19 18 19 20 18 21 24

Lotion 3 (20%) 21 21 19 0 17 25 22

Retention Lotion 1 (20%) 18 22 19 22 20 20 19

Cream 1 (20%) 22 22 21 18 18 23 22

retention lotion 2 (20%) 21 20 20 19 15 20 20

Retention lotion 3 23 20 23 20 21 23 23

Clove extract (5%) 19 19 16 0 19 0 20

Miswak extract (5%) 16 0 0 0 0 0 0

Reetha extract (5%) 0 0 0 0 0 0 0

(Table 5) contd….
Polyherbal Cream and Lotion to Treat Psoriasis Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 85

Sample PNCPL/PMOB-1 PNCPL/PMOB-2 PNCPL/PMOB-3 PNCPL/PMOB-4 PNCPL/PMOB-5 PNCPL/PMOB-6 PNCPL/PMOB-7

Walnut extract (5%) 0 0 0 0 0 0 0

Shikakai extract (5%) 0 0 17 0 0 0 0

Black pepper powder (5%) 16 0 0 0 0 0 0

Eucalyptus oil (1%) 0 23 14 0 19 16 15

Thyme oil (1%) 28 28 30 27 27 27 22

Neem seed oil (1%) 0 20 0 16 18 14 15

Coconut oil (1%) 12 17 0 0 19 14 14

Cinnamon oil (1%) 27 24 26 20 17 20 20

Jojoba oil (1%) 0 16 0 0 18 17 16

Leon peel oil (1%) 15 20 0 0 18 16 18

Lemon grass oil (1%) 20 18 22 16 17 16 16

Cream 1 extracts 26 16 18 16 0 18 0

Lotion 3 extracts 23 19 18 17 0 20 0

Retention lotion 1 extracts 18 14 12 23 0 0 14

Retention lotion 2 extracts 15 14 20 14 0 0 0

Retention lotion 1 oils 18 20 20 17 28 18 17

Cream 1 oils 25 24 23 21 22 22 0

Lotion 2 oils 24 22 25 19 16 22 21

Lotion 3 oils 22 21 17 18 25 25 0

Table 6. ZOI (mm) of samples against patient fungal sample.

Sample PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/
PMOF-2 PMOF-3 PMOF-4 PMOF-5 PMOF-6 PMOF-7 PMOF-8 PMOF-9 PMOF-10

Lotion 2 (20%) 17 19 24 13 14 12 12 12 15

Lotion 3 (20%) 18 21 27 13 14 14 12 16 21

Retention Lotion 1 (20%) 24 29 28 20 23 21 18 20 20

Cream 1 (20%) 19 25 29 19 20 16 13 21 22

Retention lotion 2 (20%) 16 18 18 17 15 19 25 17 16

Retention lotion 3 23 22 21 17 18 18 28 22 21

Clove extract (5%) 0 16 0 0 0 0 0 0 19

Miswak extract (5%) 0 0 0 0 0 0 0 0 0

Reetha extract (5%) 0 22 0 29 0 0 31 0 17

Walnut extract (5%) 0 0 0 0 0 32 0 0 0

Shikakai extract (5%) 0 21 0 19 0 0 26 0 18

Black pepper powder (5%) 0 0 0 0 0 30 0 0 0

Eucalyptus oil (1%) 0 0 0 0 0 0 0 0 0

Thyme oil (1%) 30 31 31 25 31 30 25 29 25

Neem seed oil (1%) 0 0 0 0 0 0 0 0 0

Coconut oil (1%) 0 0 15 0 18 19 0 0 0

Cinnamon oil (1%) 38 39 37 40 37 38 42 49 38

Jojoba oil (1%) 0 0 0 0 0 0 0 0 12

(Table 6) contd….
86 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 Joshi et al.

Sample PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/
PMOF-2 PMOF-3 PMOF-4 PMOF-5 PMOF-6 PMOF-7 PMOF-8 PMOF-9 PMOF-10

Leon peel oil (1%) 0 0 0 15 13 0 14 13 0

Lemon grass oil (1%) 33 52 32 32 16 32 41 27 35

Cream 1 extracts 0 22 18 0 0 0 0 0 0

Lotion 3 extracts 0 24 18 0 0 0 0 0 0

Retention lotion 1 extracts 0 0 21 16 0 16 17 0 0

Retention lotion 2 extracts 0 15 15 21 0 22 27 0 17

Cream 1 oils 27 27 27 22 29 25 22 26 23

Lotion 2 oils 33 32 31 27 16 30 32 31 21

Lotion 3 oils 20 26 23 20 27 20 22 27 19

Retention lotion 1 oils 34 34 33 28 18 33 36 35 35

Sample PNCPL/ PNCPL/ PNCPL/ NCPL/ PNCPL/ PNCPL/ PNCPL/ PNCPL/

PMOF-11 PMOF-12 PMOF-13 PMOF-A PMOF-B PMOF-D PMOF-F PMOF-CA

Lotion 2 (20%) 13 11 13 15 19 12 11 16

Lotion 3 (20%) 21 16 18 20 26 13 17 20

Retention Lotion 1 (20%) 22 22 23 24 28 23 20 26

Cream 1 (20%) 16 16 22 21 25 26 16 18

Retention lotion 2 (20%) 12 15 13 23 25 20 19 22

Retention lotion 3 23 22 22 20 22 20 22 20

Clove extract (5%) 0 17 18 0 17 0 0 18

Miswak extract (5%) 0 0 0 0 0 0 0 0

Reetha extract (5%) 0 18 0 26 0 24 29 22

Walnut extract (5%) 0 0 0 0 0 0 0 0

Shikakai extract (5%) 0 14 0 25 0 20 23 19

Black pepper powder (5%) 0 0 0 0 0 0 0 0

Eucalyptus oil (1%) 0 0 0 0 0 0 0 17

Thyme oil (1%) 29 26 25 26 25 32 24 26

Neem seed oil (1%) 0 0 0 0 0 0 0 14

Coconut oil (1%) 0 0 16 0 0 0 0 0

Cinnamon oil (1%) 40 39 40 42 40 42 42 50

Jojoba oil (1%) 0 0 0 0 0 0 0 15

Leon peel oil (1%) 0 16 0 22 0 0 28 13

Lemon grass oil (1%) 27 28 27 38 23 34 33 35

Cream 1 extracts 18 18 0 0 0 0 0 0

Lotion 3 extracts 18 18 0 0 0 0 0 0

Retention lotion 1 extracts 0 0 0 18 0 15 16 14

Retention lotion 2 extracts 0 0 0 22 0 19 21 19

Cream 1 oils 26 25 22 27 24 23 28 23

Lotion 2 oils 35 38 34 39 37 35 41 43

Lotion 3 oils 23 25 18 23 24 20 21 23

Retention lotion 1 oils 37 32 37 43 31 29 34 39

Polyherbal Cream and Lotion to Treat Psoriasis Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1 87

Fig. (5). ZOI (mm) of samples against patient bacterial strains.

Fig. (6). ZOI (mm) of samples against patient fungal strains.

88 Current Reviews in Clinical and Experimental Pharmacology, 2021, Vol. 16, No. 1
Thus, it can be concluded that among all the ingredients
screened, effective plant extracts and oils can be proposed
for the formulation of herbal cream and lotions for the treat-
ment of scalp psoriasis.
CONCLUSION
The present study shows a microbiological examination
of the lesions of psoriasis patients. A total of seven bacterial
and seventeen fungal strains were obtained. The objective of
the study was to formulate such herbal products that have
significant antimicrobial activity without causing any dam-
age to the skin or scalp. For this, we selected 6 herbal plant
extracts and 6 essential oils based on their individual activ-
ity. The oils and extracts were mixed in different proportions
using various combinations and cream and various lotions
were formulated. All products exhibited significant antimi-
crobial activity, however, cream 1 (C1), lotion 3 (L3), reten-
tion lotion 1 (L4) and retention lotion 3 (L6) showed the best
results among them. Nanocomposites were synthesized from
eucalyptus oil using zinc acetate and silver nitrate and were
used in the formulation retention lotion (L6). The composites
also showed considerable activity but this requires further
safety and efficacy study.
AUTHOR CONTRIBUTIONS
SJ conceptualised the work; PJ and UR conducted all
experiments, SJ and PJ drafted the manuscript; RBS and
DKS edited and finalised the manuscript.
ETHICAL DECLARATION
For the present study, human patients were used only to
collect the samples. Samples were collected from mid-region
of the hair, behind the ears, chest and back area of the two
patients for the culture of microorganisms from psoriatic
lesions. The whole study is based on in vitro experiments
where no patients were involved. Hence, there are no ethical
issues involved in the present study.
ETHICS APPROVAL AND CONSENT
PARTICIPATE
Not applicable.
HUMAN AND ANIMAL RIGHTS
No Animals/Humans were used for studies that are basis
of this research.
CONSENT FOR PUBLICATION
Not applicable.
AVAILABILITY OF DATA AND MATERIALS
Not applicable.
FUNDING
This work was supported by Patanjali Ayurved Ltd.
through the internal funding system.
CONFLICT OF INTEREST
The authors declare no conflict of interest, financial or
otherwise.
Joshi et al.
ACKNOWLEDGEMENTS
Declared none.
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